Pollination: self-fertilization strategy in an orchid

Mating in flowering plants normally relies on animals, wind, gravity or secretion to convey pollen grains from the male (anther) to the female (stigma) organ. Here we describe a new type of self-pollination mechanism in the tree-living orchid Holcoglossum amesianum, in which the bisexual flower turns its anther against gravity through 360 degrees in order to insert pollen into its own stigma cavity - without the aid of any pollinating agent or medium. This mode of self-pollination, which occurs under windless, drought conditions when insects are scarce, adds to the variety of mechanisms that have evolved in angiosperms to ensure their reproductive success.     

Direct ligand-receptor complex interaction controls Brassica self-incompatibility

Many higher plants have evolved self-incompatibility mechanisms to prevent self-fertilization. In Brassica self-incompatibility, recognition between pollen and the stigma is controlled by the S locus, which contains three highly polymorphic genes: S-receptor kinase (SRK), S-locus protein 11 (SP11) (also called S-locus cysteine-rich protein; SCR) and S-locus glycoprotein (SLG). SRK encodes a membrane-spanning serine/threonine kinase that determines the S-haplotype specificity of the stigma, and SP11 encodes a small cysteine-rich protein that determines the S-haplotype specificity of pollen. SP11 is localized in the pollen coat. It is thought that, during self-pollination, SP11 is secreted from the pollen coat and interacts with its cognate SRK in the papilla cell of the stigma to elicit the self-incompatibility response. SLG is a secreted stigma protein that is highly homologous to the SRK extracellular domain. Although it is not required for S-haplotype specificity of the stigma, SLG enhances the self-incompatibility response; however, how this is accomplished remains controversial. Here we show that a single form of SP11 of the S8 haplotype (S8-SP11) stabilized with four intramolecular disulphide bonds specifically binds the stigma membrane of the S8 haplotype to induce autophosphorylation of SRK8, and that SRK8 and SLG8 together form a high-affinity receptor complex for S8-SP11 on the stigma membrane.     

甘薯花粉—柱头乳突细胞间相互作用的细胞学研究

以甘薯鸟吃种和高自一号为亲本进行人工自花授粉;以甘薯鸟吃种和高自一号为母本,河北351为父本进行杂交授粉.观察了花粉在柱头上的行为,柱头乳突细胞表面糖蛋白的定位分布,授粉前后乳突细胞超微结构的变化.结果表明,不育组合首先表现出花粉在柱头上粘附和水合能力很弱,花粉不萌发.能育组合授粉后花粉在柱头上的粘附水合能力很强,多数花粉萌发.但有的花粉管不进入柱头,仅在乳突细胞表面爬行,或提前破裂,释放内容物;以及在花粉管接触的柱头乳突细胞中沉积胼胝质.糖蛋白(钉红着色物质)在能育与不育组合的柱头乳突细胞壁上的分布有明显的差异,作者用细胞化学定位法验证了糖蛋白与自交不育可能存在的相关性.鸟吃种柱头乳突细胞杂交授粉后12h其超微结构明显退化;而自花授粉后3h就已明显退化.柱头乳突细胞退化的早晚可能与所授花粉是否能育相关.     

The S-locus receptor kinase is inhibited by thioredoxins and activated by pollen coat proteins

The self-incompatibility response in Brassica allows recognition and rejection of self-pollen by the stigmatic papillae. The transmembrane S-locus receptor kinase (SRK), a member of the receptor-like kinase superfamily in plants, mediates recognition of self-pollen on the female side, whereas the S-locus cysteine-rich protein (SCR) is the male component of the self-incompatibility response. SCR is presumably located in the pollen coat, and is thought to be the SRK ligand. Although many receptor-like kinases have been isolated in plants, the mechanisms of signal transduction mediated by these molecules remain largely unknown. Here we show that SRK is phosphorylated in vivo within one hour of self-pollination. We also show that, in vitro, autophosphorylation of SRK is prevented by the stigma thioredoxin THL1 in the absence of a ligand. This inhibition is released in a haplotype-specific manner by the addition of pollen coat proteins. Our data indicate that SRK is inhibited by thioredoxins and activated by pollen coat proteins.     

油用牡丹‘凤丹’主要传粉昆虫的传粉行为比较

【目的】显花植物大多具有多种传粉者,判断最有效传粉者的直接证据是对主要传粉者的传粉效率进行研究。‘凤丹’(Paeonia ostii‘Feng Dan’)为一种新型木本食用油料植物,繁育系统为部分自交亲和、异交,需要传粉者活动才能完成授粉过程。笔者对‘凤丹’居群的访花昆虫种类及其传粉行为进行研究,旨在查清‘凤丹’主要传粉昆虫种类及其传粉效率,为提高‘凤丹’结籽率、改善种子质量、加大栽培规模提供传粉生物学参考。【方法】采用定点调查法对‘凤丹’主要传粉昆虫及其访花频率、单花停留时间、访花时柱头接触率进行观测,采用跟踪调查法对主要传粉昆虫连续两次访花的飞行距离、访固定花朵数目时的被访植株数进行观测,通过田间采集与室内镜检对主要传粉昆虫体表携粉量与单次访花后柱头花粉落置量进行统计。【结果】‘凤丹’访花昆虫主要有蜂类、蚁类、甲虫类及蝇类共4个类群,主要传粉昆虫为意大利蜜蜂(Apis mellifera)、中华蜜蜂(A. cerana)与黄熊蜂(Bombus flavescens)。意大利蜜蜂与中华蜜蜂的访花行为相似。黄熊蜂的访花频率极显著高于意大利蜜蜂和中华蜜蜂的(P<0.01),单花停留时间极显著低于意大利蜜蜂和中华蜜蜂的(P<0.01); 黄熊蜂的柱头接触率与意大利蜜蜂及中华蜜蜂的差异不显著。黄熊蜂的访花飞行距离极显著高于意大利蜜蜂和中华蜜蜂的(P<0.01),访固定花朵数目时的被访植株数极显著高于意大利蜜蜂和中华蜜蜂的(P<0.01),更能促进植株的异交授粉。黄熊蜂的体表携粉量极显著高于意大利蜜蜂和中华蜜蜂的(P<0.01),单次访花后柱头花粉落置量极显著高于意大利蜜蜂和中华蜜蜂的(P<0.01),花粉转移能力更高。【结论】黄熊蜂、意大利蜜蜂和中华蜜蜂均为‘凤丹’有效传粉昆虫,以黄熊蜂传粉效率最高,中华蜜蜂与意大利蜜蜂其次,二者具有相近传粉效率。     

药用植物丹参的传粉效率和结实率的研究

药用植物丹参Salvia miltiorrhiza Bge.在自然授粉中结实率约为80％,其花柱的授粉率较高,达到76％,为结实率的9.5倍。柱头上的平均花粉量为3－12个,但多数为来自自花的花粉,散粉期花粉活力较高,从不同处理下结实率实验看,丹参的有性繁殖主要依靠异花授粉,其自花授粉的结实率为0.6%。     

A guided tour: Pollen tube orientation in flowering plants

Sexual reproduction in flowering plants requires that two sperm cells are delivered to the embryo sac where double fertilization of an egg cell and of a central cell results in the formation of a diploid embryo and of the triploid nutritional endosperm tissue. The immobile male gametes are delivered to the im- mobile female gametophyte by a single cell, the pollen tube. The pollen tube must be able to germinate on a genetically appropriate stigma and it must be directed through the transmitting tract of the style from where it must target an ovule. Moreover, the pollen tube must enter the ovule at a defined opening, the micropyle, grow toward one of the two synergids and release the two sperm cells upon contact. This complex process requires recognition events with chemically based or physically supported cell-cell communication as well as directional cues for the growing pollen tube. A number of molecules and mechanisms have been implicated in pollen tube guidance which are summarized in this review.     

药用植物党参传粉效率和结实率的研究

药用植物党参Ｃodonopsis pilosula(Franch.)Nannf.在自然授粉中结实率约为６％,其花柱的授粉率较高的,达７５％,为结实率的１２．５倍,柱头上的平均花粉量为７－１９个,但多数为来自自花的花粉,散粉期花粉活力较高,从不同处理下结实率实验看,党参的有性繁殖主要依靠异花授粉,其自身花授粉的结实率为１．０％,作者认为,在影响结实率的诸多因素中,异花授粉的效率以及植物对异花授粉的依赖程度是重要因素。     

紫薇受粉习性及花粉管生长的研究

用脱色苯胺蓝对紫薇花粉和花粉管进行染色,在荧光显微镜下观察其自然条件下的受粉习性及花粉管生长行为,以确定花粉管通道的形成时期．结果表明,盛花期的紫薇在开花后1.5h内完成受粉,花粉落量多,受粉率为100％;开花后24h左右进行受精;单果种子数为36～50粒,认为可以用花粉管通道法转化紫薇,导入外源DNA的最佳时机为开花后24h左右.     

药用植物丹参传粉因子的研究

药用植物丹参（Salvia miltiorrhiza Bge．）在自然授粉中结实率约为8％。其花柱的授粉率较高的,可达76％,为结实率的9．5倍,柱头上的平均花粉量为3－12个,但多数为自花花粉,散粉期花粉活力较高。从不同处理下结实率实验看,丹参的有性繁殖主要依靠异花授粉,其自花授粉的结实率为0．6％。作者认为,在影响结实率的诸多因素中,异花授粉的效率以及植物对异花授粉的依赖程度是重要因素。     

不同授粉方式对祝光苹果受精过程的影响

祝光苹果自花授粉后2小时,花粉粒在柱头上萌发。授粉当日和次日,花粉管在花柱中生长。授粉后第三日,花粉管停止生长,最长花粉管平均长至花柱全长的26％。此时,花粉管先端明显膨大。而后,未见到有抵达花柱基部的花粉管,也未见到受精过程的发生。 异花授粉后4—6小时,大量花粉粒在柱头上萌发。授粉后1—2日,花粉管径直向花柱基部生长。授粉后50小时,花粉管穿过花柱基部,进入子室内。授粉后3—4日,观察到进入胚囊的花粉管将内含物释放到一个已退化的助细胞中。授粉后5—6日,观察到双受精过程。     

川党参传粉效率的研究

药用植物川党参Codonopsis tangshen Oliv.在自然授粉下结实率约为7．5％,但其花柱的授粉率高达79％,为结实率的10．5倍。柱头上的平均花娄量为5－16个,但多数为来自自花的花粉,散粉期花粉活力高,从不同处理下结实率实验看,川党参的有性繁殖主要依靠异花授粉,其自花授粉的结实率为1．7％,作者认为,在影响结实率的诸多因素中,异花授粉的效率以及植物对异花授粉的领带程度是重要因素。     

Context-dependent autonomous self-fertilization yields reproductive assurance and mixed mating

The evolution of self-fertilization in hermaphrodites is opposed by costs that decrease the value of self progeny relative to that of outcross progeny. However, self-fertilization is common in plants; 20% are highly selfing and 33% are intermediate between selfing and outcrossing. Darwin proposed an adaptive benefit of self-pollination in providing reproductive assurance when outcrossing is impossible. Moreover, if outcross pollen receipt is inconsistent within or between years, these conditions likewise favour self-pollination, and this can result in a mixture of self and outcross seed production (mixed mating). Despite wide acceptance, the reproductive assurance hypothesis has lacked the support of complete empirical evidence to show that variable pollination can create both the ecological and genetic conditions favouring self-pollination. We recently showed in Collinsia verna that during periods of infrequent pollinator visits, autonomous self-pollination boosted seed output per flower, the key ecological condition. Here we show low inbreeding depression and marker-based estimates of selfing, demonstrating that when the pollination environment in wild populations necessitates reproductive assurance, selfing rates increase. We provide a complete demonstration of reproductive assurance under variable pollination environments and mechanistically link reproductive assurance to intermediate selfing rates through mixed mating.     

开花植物延迟自交的研究进展

延迟自交是植物在单朵花开花末期主动自花授粉的一种繁殖方式。为了补充及完善植物进化和传粉生物学研究的综合数据资料,笔者概述了延迟自交的发生途径、试验研究方法、“两全其美”繁殖保障假说等方面的进展,探讨了其适应性意义,并对今后的研究方向提出了建议。目前,延迟自交现象广泛发生在43科61属74种植物中,其中,52种植物的发生途径是单一的,22种植物由两种途径共同作用。在单一途径中,柱头弯曲引起的延迟自交最为常见;不完全雌雄异熟和雌雄异位减小两种途径共同作用促进的延迟自交最为常见。对延迟自交的观察,多采用宏观传粉生物学手段,少有研究将宏观传粉生物学与遗传学手段相结合为延迟自交的发生提供最严谨的证据。延迟自交的繁殖保障假说表明,植物在优先保障异交进行的前提下,当传粉环境不可预测时,延迟自交能为植物的繁殖成功提供保障,被认为是结合了自交和异交优点的“两全其美”的交配系统。此外,对于少胚珠物种,昆虫单次授粉后即可充分授粉;为节约植物资源,还可阻止延迟自交的发生。     

井冈山地区铁皮石斛的繁殖生物学研究

通过野外实验观察和人工控制套袋等方法,从铁皮石斛的花部基本特征、开花动态、花粉活力、柱头可授性、杂交指数、套袋实验等方面对其繁殖生物学进行了研究。结果显示:(1)井冈山地区铁皮石斛4～6月开花,种群花期近60 d,多数花(50%)在约30 d内开放,果实一般需要180 d左右才能成熟。(2)铁皮石斛花粉活力开花后24 h即可达到(62.67±6.43)%,至开花第三天,活力可达到90%,柱头可授性达到最高,至第5天柱头仍保持高可授性。(3)套袋实验表明,部分自交亲和,异交,需要传粉者。自然条件下未观察到井冈山铁皮石斛居群结实,人工自花授粉率为22.2%。该研究结果为保护和利用井冈山铁皮石斛种质资源提供了科学依据。     

雌雄异型异熟青钱柳幼龄林开花习性及交配系统分析

青钱柳为典型的雌雄异型异熟树种,包括雌花先熟个体(雌先型,PG)和雄花先熟个体(雄先型, PA)两种交配型。同一交配型内雌雄花期错开而交配型间雌雄花同步的开花生物学特性可能对青钱柳种子产量和质量有重要影响。为了解青钱柳群体的交配系统,对江苏溧阳陶峰和安徽红琊山林场的7年生青钱柳人工林群体进行了连续2 a花期物候学观测,包括林分中交配型的比例、不同交配型的雌雄开花顺序、开花持续时间及花期相遇特点。结果表明:受环境条件的影响,青钱柳花期为4月下旬至5月下旬,有一定波动; 雌、雄花期持续时间分别为9～19 d(最长21 d)和2～9 d。在人工林幼龄群体中,开花植株有2类共5种表现型,其中两性植株包括雌先型(PG)、雄先型(PA)和同步型(SC),单性植株包括雌株(F)和雄株(M); 观察发现幼林群体中雌株居多,两性植株比例较小。2015年溧阳陶峰青钱柳人工林中开花率达到73.2%,两性植株占28.8%,PA和PG比例1:1.2; 而红琊山林场的开花率仅为38.6%,其中16.7%为两性植株,PG比例较高。连续2 a的定株观察还表明:开花表现型的变化主要为单性植株转变为两性植株; PA和PG表达稳定,极少发生逆转; 两性植株中,有52.2%的植株上雌、雄花期完全错开,而47.8%的植株上雌、雄花花期有部分(少量全部)重叠。相关分析表明,开花状况与母树胸高断面积显著相关,显著性大小顺序为雌先型﹥雄先型﹥雄株﹥雌株﹥未开花植株。多重比较表明:两性植株的平均胸高断面积与单性植株、未开花植株差异显著; 开花植株中雌先型、雄先型和雄株平均胸高断面积差异不显著,但三者与雌株存在显著差异。由此推测青钱柳开花与否和开花表现型明显受植株营养积累的影响。     

The S receptor kinase determines self-incompatibility in Brassica stigma

The self-incompatibility possessed by Brassica is an intraspecific reproductive barrier by which the stigma rejects self-pollen but accepts non-self-pollen for fertilization. The molecular/biochemical bases of recognition and rejection have been intensively studied. Self-incompatibility in Brassica is sporophytically controlled by the polymorphic S locus. Two tightly linked polymorphic genes at the S locus, S receptor kinase gene (SRK) and S locus glycoprotein gene (SLG), are specifically expressed in the papillar cells of the stigma, and analyses of self-compatible lines of Brassica have suggested that together they control stigma function in self-incompatibility interactions. Here we show, by transforming self-incompatible plants of Brassica rapa with an SRK28 and an SLG28 transgene separately, that expression of SRK28 alone, but not SLG28 alone, conferred the ability to reject self (S28)-pollen on the transgenic plants. We also show that the ability of SRK28 to reject S28 pollen was enhanced by SLG28. We conclude that SRK alone determines S haplotype specificity of the stigma, and that SLG acts to promote a full manifestation of the self-incompatibility response.     

Maize transformation via pollen tube pathway and the inheritance of transgene in progeny

The herbicide-resistant gene als was introduced into maize inbred line Qi319 through pollen-tube pathway. The inheritance of transgene was studied up to 33 generation. Transformation was performed by applying plasmid DNA carrying solution on the severed styles after self-pollination, or just prior to selfpoll/nation. After herbicide screening, PCR and Southern blot analysis, seventeen plants were confirmed positive in TO generation, and sixteen of them were self-bred to construct family to 33 progeny. Through PCR analysis and herbicide screening in offspring, four lines were confirmed following a 3 : 1 Mendelian segregation ratio. In other lines the numbers of positive plants were lower than expected. Therefore, it is necessary to select large population in progeny to produce stable inherited lines when transgene was delivered by pollen tube method.     

自交不亲和甘蓝的花粉萌发与花柱内保护酶活性变化

研究了柱头提取液对自交不亲和甘蓝花粉萌发、花粉管生长以及自花、异花授粉过程中花柱内过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性动态变化.结果表明:自花柱头提取液能显著抑制花粉萌发和花粉管的生长,而异花柱头提取液能提高花粉萌发率和花粉管的生长;授粉过程中花柱内SOD,POD和CAT活性都显著上升,而自花授粉上升幅度明显低于异花授粉.花柱内蛋白质含量在授粉过程中都会上升,但自花授粉在授粉后最初3～5min出现一个含量峰值.